Assay for detection of adenosine phosphorylase from mycoplasmas

Bonissol, C; Sasaki, Tsuguo; Stoiljkovic, Bob

doi:10.1016/0769-2609(88)90075-0

Cited by 8 publications

(3 citation statements)

References 9 publications

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“…Mycoplasma contamination or experimental infection of cell lines, PBMC, HIV-1 stocks, and fetal calf serum was routinely tested by four different methods: culture in SP-4 mycoplasma medium (38), fluorescence staining with 4',6diamine-2'-phenylindole (DAPI) after coculture (3 and 6 days) with 3T6 indicator cells (32), detection of adenosine phosphorylase in cell culture supernatants (5), and microbiological cultures (1). In order to avoid cross-contamination of cell cultures, manipulations were performed essentially by the method of McGarrity et al (21).…”

Section: Materuils and Methodsmentioning

confidence: 99%

Role of mycoplasma infection in the cytopathic effect induced by human immunodeficiency virus type 1 in infected cell lines

et al. 1992

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In addition to previously reported tetracycline analogs, other antibiotics known for antimycoplasmal activities inhibited the cytopathic effect in CEM c113 cells infected with human immunodeficiency virus type 1 (HIV-1) or HIV-2 but were unable to block virus replication. A contaminating mycoplasma was isolated from our CEM c113 cells and identified as a strain of Mycoplasmafermentans. Following infection of lymphoblastoid (CEM) or promonocytic (U937 and THP1) cell lines with HIV-1, cytopathic effect was observed only in association with mycoplasmal contamination. Moreover, HIV-1 infection of U937 cells after experimental inoculation with a human isolate of M. fermentans led to pronounced cell killing. We have verified that this effect is not merely an artifact caused by arginine and/or glucose depletion in the cell culture medium. These results confirm that mollicutes, in particular M. fermentans, are able to act synergistically with HIV-1 to kill infected cells in some in vitro systems.

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Section: Materuils and Methodsmentioning

confidence: 99%

Role of mycoplasma infection in the cytopathic effect induced by human immunodeficiency virus type 1 in infected cell lines

et al. 1992

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“…The assay of adenosine phosphorylase has been reported as a test to detect a contamination of cultured cells by mycoplasma [Bonissol et al, 1988]. In the LoVo cell line, however, the multiple specific assays used to detect and remove mycoplasmas revealed that, overall, a contamination can be excluded.…”

Section: Discussionmentioning

confidence: 99%

Role of the phosphorolysis of deoxyadenosine in the cytotoxic effect of the combination of deoxyadenosine and deoxycoformycin on a human colon carcinoma cell line (LoVo)

et al. 2000

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In LoVo cells, phosphorolytic activity acting on deoxyadenosine plays a major role in the resistance to the cytotoxic effect of the combination of deoxynucleoside with deoxycoformycin. In fact, the observed dependence of toxicity on cell density appears to be related to the metabolic conversion of deoxyadenosine into adenine. The phosphorylation of the deoxynucleoside, which represents the first step towards the formation of the cytotoxic agent dATP, proceeds at a significantly lower rate as compared to the phosphorolysis of deoxyadenosine. The analysis of the levels of deoxyadenosine and its derivatives in the incubation media reveals that the rates of disappearance of deoxyadenosine and of formation of adenine increase in concert with the reduction of the effect on cell survival.

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“…Purine phosphorylase and thymidine phosphorylases from mycoplasmas have been implicated in various cytogenetic effects of mycoplasmal infection of cell cultures, interfering, for example, with the incorporation of radiolabelled thymidine or of bromodeoxyuridine (16). In addition, tests based on the detection of these enzymes have been developed for the detection of mycoplasmal contamination in cell cultures (3,15). Although these two enzymes have been detected in several mycoplasmas, the plasmid pMPDEO did not hybridize (under high-stringency conditions) to genomic DNAs from other related mycoplasmas (M. genitalium, M. pneumoniae, and Mycoplasma penetrans [21) or from Mycoplasma fermentans when used as a probe in Southern blotting experiments (data not shown).…”

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confidence: 99%

Identification of Mycoplasma pirum genes involved in the salvage pathways for nucleosides

et al. 1993

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Genes encoding enzymes involved in the salvage pathway for nucleosides have been cloned and sequenced from the mollicute Mycoplasma pirum. One of them, encoding deoxyriboaldolase, was functionally identified by complementation of an Escherichia coli mutant. These genes are clustered, suggesting an operon organization, and they are immediately followed by the putative gene for the triose phosphate isomerase, an enzyme used during glycolysis.

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Assay for detection of adenosine phosphorylase from mycoplasmas

Cited by 8 publications

References 9 publications

Role of mycoplasma infection in the cytopathic effect induced by human immunodeficiency virus type 1 in infected cell lines

Role of mycoplasma infection in the cytopathic effect induced by human immunodeficiency virus type 1 in infected cell lines

Role of the phosphorolysis of deoxyadenosine in the cytotoxic effect of the combination of deoxyadenosine and deoxycoformycin on a human colon carcinoma cell line (LoVo)

Identification of Mycoplasma pirum genes involved in the salvage pathways for nucleosides

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